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Mountford, J.

Publications and source records attributed to Mountford, J..

3 recordsLinked to original sources

Enhanced short-wavelength sensitivity in the blue-tongued skink,Tiliqua rugosa

The complex visually mediated behaviors of diurnal lizards are enabled by a retina typically containing five types of opsins with the potential for tetrachromatic color vision. Despite lizards using a wide range of color signals, the limited variation in photoreceptor spectral sensitivities across lizards suggests only weak selection for species-specific, spectral tuning of photoreceptors. Some species, however, have enhanced short wavelength sensitivity, which likely helps with the detection of signals rich in ultraviolet and short wavelengths. In this study, we examined the visual system of Tiliqua rugosa, which has a UV/blue tongue, to determine the spectral sensitivity of the eye and to gain insight into this species visual ecology. Electroretinograms coupled with spectral stimulation showed peak sensitivity at 560 nm with high similarity to other lizards at wavelengths greater than 530 nm. However, at shorter wavelengths, sensitivity is enhanced leading to a spectral sensitivity curve that is 28 nm broader (full width at half height) than other lizards studied so far. The width of the curve is partially explained by a population of photoreceptors that respond more strongly to low temporal frequencies with possible peaks in sensitivity between 460 and 470 nm suggesting that they are SWS2 photoreceptors. The lack of a peak in sensitivity at 360 nm at low temporal frequencies and under a monochromatic light that suppresses the response of LWS photoreceptors, suggests that the SWS1 photoreceptors are red-shifted. In addition, the yellow and green oil droplets that are common in other diurnal lizards appear to be missing and instead, only transparent and pale-yellow oil droplets are present. LWS photoreceptors are likely paired with pale-yellow oil droplets to produce LWS photoreceptors with wider spectral sensitivity curves than in other lizards. Opsin sequencing reveals SWS1, SWS2, RH1, RH2 and LWS opsin genes that are very similar to the visual opsins detected in the green anole, Anolis carolinensis, suggesting there is little change in the spectral sensitivity of photoreceptors compared to other diurnal lizards. Since we only obtained a partial sequence of the SWS1 opsin, we were unable to determine whether amino acid substitution at tuning sites could have played a role in red-shifting the SWS1 photoreceptor spectral sensitivity. Photoreceptor densities are typically higher in central and ventral retinal regions than in the dorsal retina suggesting that higher spatial sampling is necessary at eye level and above the animal than on the ground. However, the SWS1 photoreceptors do not follow this pattern potentially due to their low abundance making them less relevant to high acuity visual tasks. Our findings demonstrate that there are possibly multiple mechanisms acting synergistically in the visual system of T. rugosa to enhance short wavelength sensitivity between 360 and 530 nm. While it is tempting to suggest that this is an adaptation to facilitate the detection of the blue tongues of conspecifics, additional experiments are necessary to determine its ecological relevance. Summary statementColor vision and the distribution of photoreceptor subtypes in T. rugosa

neuroscience↗

In vitro and In vivo Genetic Disease Modelling via NHEJ precise deletions using CRISPR/Cas9

Development of advanced gene and cell therapies for the treatment of genetic diseases requires confident animal and cellular models to test their efficacy and is crucial in the cases where no primary samples from patients are available. CRISPR/Cas9 technology, has become one of the most spread endonuclease tools for editing the genome at will. Moreover, it is known that the use of two guides tends to produce the precise deletion between the guides via NHEJ. Different distances between guides were tested (from 8 to 500 base pairs). We found that distances between the two cutting sites and orientation of Cas9 protein-DNA interaction are important for the efficiency within the optimal range (30-60 bp), we could obtain new genetically reproducible models for two rare disease, a Pyruvate Kinase Deficiency model, using human primary cells, and a (for in vivo primary hyperoxaluria therapeutic mouse model. We demonstrate that the use of a 2-guideRNAs at the optimal distance and orientation is a powerful strategy for disease modelling in those diseases where the availability of primary cells is limited.

bioengineering↗

Nanotopography reveals metabolites that maintain the immunosuppressive phenotype of mesenchymal stem cells

Mesenchymal stem cells (MSCs) are multipotent stem cells that are immunosuppressive and thus of considerable therapeutic potential in transplant operations. However, MSCs rapidly differentiate once in culture, making their large-scale expansion for use in immunosuppressive therapies challenging. Although the differentiation mechanisms of MSCs have been extensively investigated using materials, little is known about how materials can modulate paracrine activities of MSCs. Here, we show for the first time that nanotopography can control the immunomodulatory capacity of MSCs through decreased intracellular tension increasing oxidative glycolysis. We also use the nanotopography to identify bioactive metabolites that modulate intracellular tension, growth and immunomodulatory phenotype of MSCs in standard culture. Our findings show a novel route to support large-scale expansion of functional MSCs for therapeutic purposes

bioengineering↗